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At least 199 records · Page 11

All sources NASA NTRS DOE OSTI

DOE OSTI2022

X-ray fluorescence microscopy methods for biological tissues

Abstract Synchrotron-based X-ray fluorescence microscopy is a flexible tool for identifying the distribution of trace elements in biological specimens across a broad range of sample sizes. The technique is not particularly limited by sample type and can be performed on ancient fossils, fixed or fresh tissue specimens, and in some cases even live tissue and live cells can be studied. The technique can also be expanded to provide chemical specificity to elemental maps, either at individual points of interest in a map or across a large field of view. While virtually any sample type can be characterized with X-ray fluorescence microscopy, common biological sample preparation methods (often borrowed from other fields, such as histology) can lead to unforeseen pitfalls, resulting in altered element distributions and concentrations. A general overview of sample preparation and data-acquisition methods for X-ray fluorescence microscopy is presented, along with outlining the general approach for applying this technique to a new field of investigation for prospective new users. Considerations for improving data acquisition and quality are reviewed as well as the effects of sample preparation, with a particular focus on soft tissues. The effects of common sample pretreatment steps as well as the underlying factors that govern which, and to what extent, specific elements are likely to be altered are reviewed along with common artifacts observed in X-ray fluorescence microscopy data.

Pushie, M. Jake (ORCID:0000000174945427)↗

DOE OSTI2024

X-ray fluorescence mapping of brain tissue reveals the profound extent of trace element dysregulation in stroke pathophysiology

Abstract The brain is a privileged organ with regard to its trace element composition and maintains a robust barrier system to sequester this specialized environment from the rest of the body and the vascular system. Stroke is caused by loss of adequate blood flow to a region of the brain. Without adequate blood flow ischaemic changes begin almost immediately, triggering an ischaemic cascade, characterized by ion dysregulation, loss of function, oxidative damage, cellular degradation, and breakdown of the barrier that helps maintain this environment. Ion dysregulation is a hallmark of stroke pathophysiology and we observe that most elements in the brain are dysregulated after stroke. X-ray fluorescence-based detection of physiological changes in the neurometallome after stroke reveals profound ion dysregulation within the lesion and surrounding tissue. Not only are most elements significantly dysregulated after stroke, but the level of dysregulation cannot be predicted from a cell-level description of dysregulation. X-ray fluorescence imaging reveals that the stroke lesion retains <25% of essential K+ after stroke, but this element is not concomitantly elevated elsewhere in the organ. Moreover, elements like Na+, Ca2+, and Cl− are vastly elevated above levels available in normal brain tissue (>400%, >200%, and >150%, respectively). We hypothesize that weakening of the blood–brain barrier after stroke allows elements to freely diffuse down their concentration gradient so that the stroke lesion is in equilibrium with blood (and the compartments containing brain interstitial fluid and cerebrospinal fluid). The change observed for the neurometallome likely has consequences for the potential to rescue infarcted tissue, but also presents specific targets for treatment.

Biochemistry & Molecular Biology↗

DOE OSTI2020

Skeletal tissue regulation by catalase overexpression in mitochondria

Accumulation of oxidative damage from excess reactive oxygen species (ROS) may contribute to skeletal aging and mediate adverse responses to physiological challenges. Wild-type (WT) mice and transgenic mice (male, 16 wk of age) with human catalase targeted to the mitochondria (mCAT) were analyzed for skeletal responses to the remodeling stimuli of combined hind-limb unloading and exposure to ionizing radiation ( 137 Cs, 2 Gy). Treatment for 2 wk caused lipid peroxidation in the bones WT but not mCAT mice, showing that transgene expression mitigated oxidative stress. Ex vivo osteoblast colony growth rate was 95% greater in mCAT than WT mice and correlated with catalase activity levels ( P < 0.005, r = 0.67), although terminal osteoblast and osteoclast differentiation were unaffected. mCAT mice had lower cancellous bone volume and cortical size than WT mice. Ambulatory control mCAT animals also displayed reduced cancellous and cortical structural properties compared with control WT mice. In mCAT but not WT mice, treatment caused an unexpectedly rapid radial expansion (+8% cortical area, +22% moment of inertia), reminiscent of compensatory bone growth during advancing age. In contrast, treatment caused similar structural deficits in cancellous tissue of mCAT and WT mice. In sum, mitochondrial ROS signaling via H 2 O 2 was important for the acquisition of adult bone structure and catalase overexpression failed to protect cancellous tissue from treatment. In contrast, catabolic stimuli caused radial expansion in mCAT not WT mice, suggesting that mitochondrial ROS in skeletal cells act to suppress tissue turnover in response to remodeling challenges.

Cell Biology↗

DOE OSTI2020

Lawrence Livermore National Laboratory ABSL-3 Tissue Digester Run Data Report

Purpose: Provide data from a water run of the LLNL tissue digestor to confirm operational status for possible extension of Medical Waste permit, since the annual recertification maintenance and testing could not take place due to the shelter-in-place of the tissue digester vendor who is located outside of California. This is a unique medical waste device and the vendor who certifies our other medical waste equipment (permitted autoclaves), does not service the tissue digester.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Lawrence Livermore National Laboratory ABSL-3 Tissue Digester 12 June 2020 Run Data Report, Report 2

Purpose: Provide data from a water run of the LLNL tissue digester to confirm operational status for possible extension of Medical Waste permit, since the annual recertification maintenance and testing could not take place due to the shelter-in-place of the tissue digester vendor who is located outside of California. This is a unique medical waste device and the vendor who certifies our other medical waste equipment (permitted autoclaves), does not service the tissue digester.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Lawrence Livermore National Laboratory ABSL-3 Tissue Digester 12 June 2020 Run Data Report

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Lawrence Livermore National Laboratory ABSL-3 Tissue Digester (8 July 2020 Run Data Report, Report 3)

42 ENGINEERING↗

DOE OSTI2023

Non-Equilibrium Plasma Interactions with Biomaterials, Biological Solutions and Tissues

Cold atmospheric pressure plasma discharges offer an abundant source of reactive oxygen and nitrogen species (RONS) at room temperature enabling unique interactions with biomaterials, biological solutions and tissues. These interactions particularly with living matter are presently an important intellectual frontier in plasma science with promising potential applications ranging from human health care to advanced biomaterial processing. Exciting case studies have been reported that illustrate the huge potential of cold atmospheric plasma technology in wound healing and cancer treatment. The interaction of plasma with conducting and dielectric biomaterials such as tissue strongly influences the plasma properties. In turn this changes the impact of the plasma on the biomaterial. Particularly in the case of living matter, liquid based solutions are ubiquitous which complicates interfacial processes. The lack of insight into the underlying mechanisms of the interaction of plasma with wounds and tumors is currently a bottleneck for the further development of the technology and gives rise to many interesting scientific questions. This project was focused on plasma properties and kinetics during plasma-biomaterial interactions. Both DC pulsed and RF driven atmospheric pressure plasma jets, extensively used by the plasma community were studied. The bio-interfaces included hydrogel as a tissue model, (saline) solutions, bacteria and virus. Plasma diagnostics used include Thomson scattering, Rayleigh scattering, Raman scattering, (two-photon absorption) laser induced fluorescence, optical emission spectroscopy, absorption spectroscopy, molecular beam mass spectrometry and fast imaging allowing to determine electron densities and temperatures, ionic species, reactive species including radicals, gas temperatures, gas composition, electric fields and solution components transferred to the gas phase.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

DOE OSTI2023

Differential transcriptomic alterations in nasal versus lung tissue of acrolein-exposed rats

Introduction: Acrolein is a significant component of anthropogenic and wildfire emissions, as well as cigarette smoke. Although acrolein primarily deposits in the upper respiratory tract upon inhalation, patterns of site-specific injury in nasal versus pulmonary tissues are not well characterized. This assessment is critical in the design of in vitro and in vivo studies performed for assessing health risk of irritant air pollutants. Methods: In this study, male and female Wistar-Kyoto rats were exposed nose-only to air or acrolein. Rats in the acrolein exposure group were exposed to incremental concentrations of acrolein (0, 0.1, 0.316, 1 ppm) for the first 30 min, followed by a 3.5 h exposure at 3.16 ppm. In the first cohort of male and female rats, nasal and bronchoalveolar lavage fluids were analyzed for markers of inflammation, and in a second cohort of males, nasal airway and left lung tissues were used for mRNA sequencing. Results: Protein leakage in nasal airways of acrolein-exposed rats was similar in both sexes; however, inflammatory cells and cytokine increases were more pronounced in males when compared to females. No consistent changes were noted in bronchoalveolar lavage fluid of males or females except for increases in total cells and IL-6. Acrolein-exposed male rats had 452 differentially expressed genes (DEGs) in nasal tissue versus only 95 in the lung. Pathway analysis of DEGs in the nose indicated acute phase response signaling, Nrf2-mediated oxidative stress, unfolded protein response, and other inflammatory pathways, whereas in the lung, xenobiotic metabolism pathways were changed. Genes associated with glucocorticoid and GPCR signaling were also changed in the nose but not in the lung. Discussion: These data provide insights into inhaled acrolein-mediated sex-specific injury/inflammation in the nasal and pulmonary airways. The transcriptional response in the nose reflects acrolein-induced acute oxidative and cytokine signaling changes, which might have implications for upper airway inflammatory disease susceptibility.

Alewel, Devin I.↗

DOE OSTI2024

Analysis of 26S Proteasome Activity across Arabidopsis Tissues

Plants utilize the ubiquitin proteasome system (UPS) to orchestrate numerous essential cellular processes, including the rapid responses required to cope with abiotic and biotic stresses. The 26S proteasome serves as the central catalytic component of the UPS that allows for the proteolytic degradation of ubiquitin-conjugated proteins in a highly specific manner. Despite the increasing number of studies employing cell-free degradation assays to dissect the pathways and target substrates of the UPS, the precise extraction methods of highly potent tissues remain unexplored. Here, we utilize a fluorogenic reporting assay using two extraction methods to survey proteasomal activity in different Arabidopsis thaliana tissues. This study provides new insights into the enrichment of activity and varied presence of proteasomes in specific plant tissues.

26S proteasome↗

DOE OSTI2021

Toward Revealing Microcystin Distribution in Mouse Liver Tissue Using MALDI-MS Imaging

Cyanotoxins can be found in water and air during cyanobacterial harmful algal blooms (cHABs) in lakes and rivers. Therefore, it is very important to monitor their potential uptake by animals and humans as well as their health effects and distribution in affected organs. Herein, the distribution of hepatotoxic peptide microcystin-LR (MC-LR) is investigated in liver tissues of mice gavaged with this most common MC congener. Preliminary matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) imaging experiments performed using a non-automated MALDI matrix deposition device and a MALDI-time-of-flight (TOF) mass spectrometer yielded ambiguous results in terms of MC-LR distribution in liver samples obtained from MC-LR-gavaged mice. The tissue preparation for MALDI-MS imaging was improved by using an automated sprayer for matrix deposition, and liver sections were imaged using an Nd:YAG MALDI laser coupled to a 15 Tesla Fourier-transform ion cyclotron resonance (FT-ICR)-mass spectrometer. MALDI-FT-ICR-MS imaging provided unambiguous detection of protonated MC-LR (calculated m/z 995.5560, z = +1) and the sodium adduct of MC-LR (m/z 1017.5380, z = +1) in liver sections from gavaged mice with great mass accuracy and ultra-high mass resolution. Since both covalently bound and free MC-LR can be found in liver of mice exposed to this toxin, the present results indicate that the distribution of free microcystins in tissue sections from affected organs, such as liver, can be monitored with high-resolution MALDI-MS imaging.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1971

Steady-state temperature distribution in living tissue modeled as cylindrical shells.

Closed form, analytical solutions to the problem of steady-state heat transfer in living tissue modeled as cylindrical shells are presented and discussed. These solutions are particularly useful for the study of temperature distributions in the extremities. Metabolic heat generation, conduction, and heat transported by the blood perfusing the tissue are considered in the model. The results demonstrate the important role that the blood stream plays in the transfer of heat inside living tissue. Solutions are also presented for the limiting cases of diminishing blood flow that would occur during vasoconstriction or occlusion of blood by external means.

Shitzer, A.↗

NASA NTRS1971

Analytical solutions to the problem of transient heat transfer in living tissue.

An analytical model of transient heat transfer in living biological tissue is considered. The model includes storage, generation, conduction, and convective transport of heat in the tissue. Solutions for rectangular and cylindrical coordinates are presented and discussed. Transient times for reaching the ?locally fully developed' temperature profile were found to be of the order of 5 to 25 min. These transients are dominated by a geometrical parameters and, to a lesser extent, by a parameter representing the ratio of heat supplied by blood flow to heat conducted in the tissue.

Shitzer, A.↗

NASA NTRS1973

Effect of lunar materials on plant tissue culture.

Lunar material collected during the Apollo 11, 12, 14, and 15 missions has been used to treat 12 species of higher plant tissue cultures. Biochemical and morphological studies have been conducted on several of these species. Tobacco tissue cultures treated with 0.22 g of lunar material exhibited increased greening more complex chloroplasts, less cytoplasmic vacuolation and greater vesiculation. Pine tissue cultures reacted to treatment by an increased deposition of tannin-like materials. The percentage of dry weight and soluble protein was increased in cultures treated with either lunar or terrestrial rock materials.

Walkinshaw, C. H.↗

NASA NTRS1973

Gas chromatography-mass spectrometry study of sterols from Pinus elliotti tissues.

A comparative study of the sterol components of slash pine (Pinus elliotti) callus tissue cultures, seeds, and seedlings was carried out using GC-MS techniques. Cholesterol, desmosterol, campesterol, stigmasterol, sitosterol and cycloeucalenol were identified in all tissues while lophenol and 24-methylenelophenol were identified in only the seed and seedlings. 24-Ethylidenelophenol was detected in trace concentrations in only the seedlings. Sitosterol was the predominant sterol component, i.e., 80.8, 38.1 and 47.8% of the tissue culture, seed and seedling sterols, respectively.

Laseter, J. L.↗

NASA NTRS1973

Intermediary metabolism during brief and prolonged low tissue temperature

The intermediary metabolism of the depressed metabolic state in the hypothermic hamster and the hibernating ground squirrel was studied by observing acetate and palmitic acid metabolisms in their tissues. The oxidative metabolism seemed to be dominant in the depressed state although synthetic reactions such as fat synthesis proceeded in some cases at a faster rate than normothermic metabolism for the same tissues. Fat syntheses proceeded in all tissues with brown fat and liver especially active. Enzymes for the synthesis of cholesterol seemed to be more temperature sensitive than enzymes for fatty acid synthesis. It was concluded that there are no great differences between metabolisms in hypothermic and hibernating animals.

Enteman, C.↗

NASA NTRS1974

Study of a number of biochemical indices of the blood and tissue of dogs after prolonged gamma-radiation

The glucose content in blood and the lipid content in serum and tissues of dogs exposed to chronic radiation for 3 and 5 years were studied. In tissues of these animals, the concentration of soluble DNA and DNA contained in DNP was studied in the spleen, lymph node (deep cervical node) and bone marrow of thigh bones. Results indicate that chronic gamma irradiation significantly changes concentrations of glucose in the blood, and that of several lipids in serum and tissues. A reduction in the concentration of DNP in tested organs reflects changes in the relative number of cells with various nuclear cytoplasmic ratios; most pronounced changes in biochemical indices occur in dogs exposed to chronic gamma radiation in doses of 125 rad per year.

Alers, I.↗

NASA NTRS1980

State of the mineral component of rat bone tissue during hypokinesia and the recovery period

Experiments were conducted on young growing rats. Hypokinesia lasting from 20 to 200 days caused retarded gain in weight and volume of the femur and delayed development of the cortical layer of the diaphysis. In contrast, the density of the cortical layer of the femoral diaphysis increased due to elevation of the mineral saturation of the bone tissue microstructures. Incorporation of Ca into the bone tissue in hypokinesia had a tendency to reduce. Partial normalization of the bone tissue mineral component occurred during a 20 day recovery period following hypokinesia.

Volozhin, A. I.↗